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. 2021 Aug 3;53:101313. doi: 10.1016/j.molmet.2021.101313

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© 2021 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Supplemental Figure 2 — Differentiation of NEUROG3-HA-P2A-Venus hiPSCs line to pancreatic endocrine progenitors (PEP) and FACS-sorting of Venus+ cells. (A–B) Immunofluorescence staining for NEUROG3, Venus, and PDX1 (A) or HA (B) at PEP stage (day 13 of differentiation). DNA counterstained with Dapi is shown as an inset within the merged images. Scale bar = 50 μM. Arrows point to a selection of cells co-expressing (A) NEUROG3, Venus, and PDX1 or (B) NEUROG3, Venus, and HA. (C) Representative flow cytometry plots showing the percentage of cells expressing NEUROG3-3HA (anti-HA antibody), Venus (anti-GFP antibody), and PDX1 (anti-PDX1 antibody) at day 13 for the differentiated SB AD3.1 and NEUROG3-HA-P2A-Venus hiPSC lines. (D) Representative flow cytometry plot showing the sorted living Venus+ cells at day 13 used for CUT&RUN experiments.